Study on the antineoplastic and toxicological effects of pomegranate (Punica granatum L.) leaf infusion using the K14-HPV16 transgenic mouse model.

Punica granatum L. (pomegranate) has been used in functional foods due to its various health benefits. However, the in vivo biological potential of its leaf remains little known. This study has aimed to characterize the antineoplastic and toxicological properties of using pomegranate leaf infusion (PLI) on transgenic mice carrying human papillomavirus (HPV) type 16 oncogenes. Thirty-eight mice were divided into 3 wild-type (WT) and 3 transgenic (HPV) groups, with exposure to 0.5% PLI, 1.0% PLI, and water. The animals' body weight, drink and food consumption were recorded. Internal organs, skin samples and intracardiac blood were collected to evaluate toxicological parameters, neoplastic lesions and oxidative stress. The results indicated that PLI was safe as no mortality, no behavioural disorders and no significant differences in the levels of microhematocrit, serum biochemical markers, internal organ histology, and oxidative stress was found among the WT groups. Histological analysis revealed that HPV animals that consumed PLI exhibited reduced hepatic, renal and cutaneous lesions compared with the HPV control group. Low-dose PLI consumption significantly diminished renal hydronephrosis lesions and relieved dysplasia and carcinoma lesions in the chest skin. Oxidative stress analysis showed that low-dose PLI consumption may have more benefits than high-dose PLI. These results suggest that oral administration of PLI has the potential to alleviate non-neoplastic and neoplastic lesions against HPV16-induced organ and skin injuries, though this requires further scientific research studies.

Mitigation of nicotine-induced developmental effects by 24-epibrassinolide in zebrafish.

Nicotine is a highly addictive substance that can cause teratogenic impacts in the embryo through redox-dependent pathways. As antioxidants, naturally occurring chemicals can protect cells from redox imbalance. The purpose of this study was to evaluate the effectiveness of 24-epibrassinolide (24-EPI), a natural brassinosteroid with well-known antioxidant properties, in protecting zebrafish embryos against nicotine's teratogenic effects. For 96 h, embryos (2 h post-fertilization - hpf) were exposed to 100 µM nicotine, co-exposed with 24-EPI (0.01, 0.1, and 1 µM), and 24-EPI alone (1 µM). Lethal and sublethal developmental characteristics were evaluated during exposure. Biochemical tests were performed at the conclusion of the exposure, and distinct behavioural paradigms were analysed 24 h later. Nicotine exposure resulted in a higher proportion of larvae with deformities, which were decreased following co-exposure to 24-EPI. Nicotine exposure also caused an increase in oxidative stress as observed by the increased activity of superoxide dismutase and catalase accompanied by an increase in the malondialdehyde levels. Besides, metabolic changes were noticed as observed by the increased lactate dehydrogenase activity that were hypothesised to be associated to nicotine-induced hypoxia which may be responsible for the increased oxidative damage. In addition, locomotor deficits were observed as well as a decrease in the acetylcholinesterase activity denoting nicotine-induced cognitive dysfunction. However, co-exposure to 24-EPI alleviated behavioural deficits and improved nicotine-induced emotional states. Overall, and although further studies are required to clarify these effects, 24-EPI showed promising ameliorative properties against the teratogenic effects induced by nicotine.

Beneficial Effects of Broccoli (Brassica oleracea var italica) By-products in Diet-induced Obese Mice.

BACKGROUND/AIM: Obesity currently affects the whole world, with greater incidence in high-income countries, with vast economic and social costs. Broccoli harvest generates many by-products equally rich in bioactive compounds with potential anti-obesity effects. This study aimed to evaluate the anti-obesity effects of broccoli by-products flour (BF) in obese mice. MATERIALS AND METHODS: A commercial high-fat diet formulation (representing a Western diet) was used to induce obesity in mice. BF (0.67% or 1.34% weight/weight) was incorporated as a chemoprevention compound into a control and a hypercholesterolemic diet, at two different concentrations, and fed for 14 weeks to C57BL/6J mice. For a therapeutic approach, two groups were fed with the hypercholesterolemic diet for 10 weeks, and then fed with BF-supplemented diets in the last 4 weeks of the study. RESULTS: BF supplementation helped to maintain a lower body weight, reduced adipose tissue accumulation, and enhanced the basal activity of superoxide dismutase and glutathione S-transferase. Although BF supplementation tended to reduce the relative liver weight increased by the Western diet, the differences were not significant. CONCLUSION: BF appears to have a beneficial effect in preventing weight gain and fat accumulation induced by hypercholesterolemic diets.

Evaluation of the developmental effects of a glyphosate-based herbicide complexed with copper, zinc, and manganese metals in zebrafish.

The use of glyphosate-based herbicides (GBH) has increased dramatically, being currently the most used herbicides worldwide. Glyphosate acts as a chelating agent, capable of chelate metals. The synergistic effects of metals and agrochemicals may pose an environmental problem as they have been shown to induce neurological abnormalities and behavioural changes in aquatic species. However, as their ecotoxicity effects are poorly understood, evaluating the impacts of GBH complexed with metals is an ecological priority. The main objective of the study was to evaluate the potentially toxic effects caused by exposure to a GBH (1 µg a.i. mL-1), alone or complexed with metals (Copper, Manganese, and Zinc (100 µg L-1)), at environmentally relevant concentrations, during the early period of zebrafish (Danio rerio) embryo development (96 h post-fertilization), a promising model for in vivo developmental studies. To clarify the mechanisms of toxicity involved, lethal and sublethal development endpoints were assessed. At the end of the exposure, biochemical and cell death parameters were evaluated and, 24 h later, different behavioural responses were assessed. The results showed that metals induced higher levels of toxicity. Copper caused high mortality, low hatching, malformations, and changes in biochemical parameters, such as decreased Catalase (CAT) activity, increased Glutathione Peroxidase (GPx), Glutathione S-Transferase (GST), reduced Glutathione (GSH) and decreased Acetylcholinesterase (AChE) activity, also inducing apoptosis and changes in larval behaviour. Manganese increased the activity of SODs enzymes. Zinc increased mortality, reactive oxygen species (ROS) levels, superoxide dismutase activity (SODs) and caused a decrease in AChE activity. Embryos/larvae exposed to the combination of GBH/Metal also showed teratogenic effects during their development but in smaller proportions than the metal alone. Although more studies are needed, the results suggest that GBH may interfere with the mechanisms of metal toxicity at the biochemical, physiological, and behavioural levels of zebrafish.

Toxicological and anti-tumor effects of a linden extract (Tilia platyphyllos Scop.) in a HPV16-transgenic mouse model.

Tilia platyphyllos Scop. is a popular broad-leaved tree, native to Central and Southern Europe. Hydroethanolic extracts rich in phenolic compounds obtained from T. platyphyllos Scop. have shown in vitro antioxidant, anti-inflammatory and antitumor properties. The aim of this work was to evaluate the therapeutic properties of a hydroethanolic extract obtained from T. platyphyllos in HPV16-transgenic mice. The animals were divided into eight groups according to their sex and phenotype. Four groups of female: HPV+ exposed to linden (HPV linden; n = 6), HPV+ (HPV water; n = 4), HPV- exposed to linden (WT linden; n = 5) and HPV- (WT water; n = 4) and four groups of male: HPV+ exposed to linden (HPV linden; n = 5), HPV+ (HPV water; n = 5), HPV- exposed to linden (WT linden; n = 5) and HPV- (WT water; n = 7). The linden (Tilia platyphyllos Scop.) extract was orally administered at a dose of 4.5 mg/10 mL per animal (dissolved in water) and changed daily for 33 days. The hydroethanolic extract of T. platyphyllos consisted of protocatechuic acid and (-)-epicatechin as the most abundant phenolic acid and flavonoid, respectively, and was found to be stable during the studied period. In two male groups a significant positive weight gain was observed but without association with the linden extract. Histological, biochemical, and oxidative stress analyses for the evaluation of kidney and liver damage support the hypothesis that the linden extract is safe and well-tolerated under the present experimental conditions. Skin histopathology does not demonstrate the chemopreventive effect of the linden extract against HPV16-induced lesions. The linden extract has revealed a favourable toxicological profile; however, additional studies are required to determine the chemopreventive potential of the linden extract.

Exposure to aluminium causes behavioural alterations and oxidative stress in the brain of adult zebrafish.

Aluminium (Al) water pollution is an increasing environmental problem. Accordingly, this study aimed to find out more about its toxic effects on aquatic organisms. Adult zebrafish were exposed to 11 mg/L of Al and the behavioural responses and its correlation with brain oxidative stress, antioxidant-defences, changes in metabolism and neurotransmission were assessed at 10, 15 and 20 days of exposure. The behavioural and locomotory responses, suggest an increase in the anxiety state, especially observed in animals exposed to Al for 15 days. The reactive oxygen species increased in a time-dependent trend, while the oxidative damage varied over exposure time. The activity of antioxidant enzymes, as superoxide dismutase, glutathione peroxidase and glutathione S-transferases, and the metallothioneins levels increased after short-term exposures and tended to decrease or stabilize at longer times. The results contribute to understand the toxic mechanisms activated by Al highlighting correlations like behavioural disorders and oxidative state.

24-Epibrassinolide protects against ethanol-induced behavioural teratogenesis in zebrafish embryo.

Embryonic studies have demonstrated the neurotoxic, teratogenic, and neurobehavioral toxicity of ethanol (EtOH). Although multiple mechanisms may contribute to these effects, oxidative stress has been described as the major damage pathway. In this regard, natural antioxidants have the potential to counteract oxidative stress-induced cellular damage. Therefore, the present study aimed to investigate the potential protective role of 24-epibrassinolide (24-EPI), a natural brassinosteroid with proved antioxidant properties, in EtOH-induced teratogenic effects during early zebrafish development. Embryos (~2 h post-fertilization - hpf) were exposed to 1 % EtOH, co-exposed to 24-EPI (0.01, 0.1 and 1 µM) and to 24-EPI alone (1 µM) for 24 h. Following exposure, biochemical evaluations were made at 26 hpf, developmental analysis was made throughout the embryo-larval period, and behavioural responses were evaluated at 120 hpf. Exposure to 1 % EtOH caused an increase in the number of malformations, which were diminished by 24-EPI. In addition, EtOH induced an accumulation of GSSG and consequent reduction of GSH:GSSG ratio, indicating the involvement of oxidative mechanisms in the EtOH-induced effects. These were reverted by 24-EPI as proved by the GSSG levels and GSH:GSSG ratio that returned to control values. Furthermore, exposure to EtOH resulted in behavioural deficits at 120 hpf as observed by the disrupted response to an aversive stimulus, suggesting the involvement of neurotoxic mechanisms. 24-EPI restored the behavioural deficits observed in a dose-dependent manner. The absence of effects in the embryos exposed solely to 24-EPI showed its safety during the exposure period. In conclusion, EtOH caused developmental teratogenicity and behavioural toxicity by inducing glutathione changes, which were prevented by 24-EPI.

Polyphenol composition and biological activity of Thymus citriodorus and Thymus vulgaris: Comparison with endemic Iberian Thymus species.

The polyphenol compositions of Thymus × citriodorus and Thymus vulgaris extracts as obtained by exhaustive hydroethanolic (HE) extraction and aqueous decoction (AD) were compared. In addition, their compositions and bioactivities were compared to those of Thymus pulegioides and Thymus mastichina, grown under the same edaphoclimatic conditions, and Thymus carnosus. Rosmarinic acid was the most abundant polyphenol followed by luteolin-hexuronide, salvianolic acids I and K. Cluster analysis suggests a similarity of the polyphenol composition of T. citriodorus and T. vulgaris. A significant antioxidant activity was observed and correlated with their polyphenol levels. The same being observed for the higher anti-proliferative activity/cytotoxicity of HE extracts on Caco-2 and HepG2 cells as compared to AD extracts. Significant association between the total phenolic compounds with the anti-proliferative activity, for both cell lines, was observed. These results support the importance of salvianolic acids levels in Thymus extracts and their in vitro anti-proliferative/cytotoxic activities.

Thymus pulegioides L. as a rich source of antioxidant, anti-proliferative and neuroprotective phenolic compounds.

The potential of Thymus pulegioides L. as an alternative and valuable source of functional ingredients has been assessed. For this purpose, the phenolic constituent profiles and the antioxidant, anti-proliferative, neuroprotective, anti-aging and anti-diabetic activities of both T. pulegioides aqueous decoctions (AD) and hydro-ethanolic extracts (HE) were studied and compared for the first time. Rosmarinic acid was the main phenolic compound, accounting for 35.2% or 47.8% of total identified phenols in AD or HE, respectively (58.5 and 151.9 mg g-1 extract). Furthermore, large amounts of luteolin-O-hexuronide (AD: 39.9 ± 2.5 mg g-1, HE: 60.8 ± 7.0 mg g-1), eriodictyol-O-hexuronide (AD: 19.9 ± 2.5 mg g-1, HE: 26.8 ± 7.0 mg g-1) and chrysoeriol hexoside (AD: 23.5 ± 0.7 mg g-1, HE: 16.0 ± 0.7 mg g-1) were found. Both extracts showed significant in vitro antioxidant activity and anti-proliferative activity against Caco-2 cells (IC50 82.25 µg mL-1 and 105.44 µg mL-1, AD and HE, respectively), and reduced hepatotoxicity (HepG2 cells). In general, both T. pulegioides extracts showed poor anti-diabetic activity, moderate anti-aging effects and high neuroprotective activity with both AD and HE extracts, at 0.5 mg mL-1, showing 80% inhibition of the acetylcholinesterase activity and 94% inhibition of the tyrosinase activity. The present study highlights the important potential of this herb as a functional food ingredient.

Ketamine induction of p53-dependent apoptosis and oxidative stress in zebrafish (Danio rerio) embryos.

Ketamine is a widely used pharmaceutical that has been detected in water sources worldwide. Zebrafish embryos were used in this study to investigate the oxidative stress and apoptotic signals following a 24h exposure to different ketamine concentrations (0, 50, 70 and 90 mg L-1). Early blastula embryos (∼2 h post fertilisation-hpf) were exposed for 24 h and analysed at 8 and 26 hpf. Reactive oxygen species and apoptotic cells were identified in vivo, at 26 hpf. Enzymatic activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE)), glutathione levels (oxidised (GSSG) and reduced (GSH)), oxidative damage (lipid peroxidation (LPO) and protein carbonyls (CO)) as well as oxidative stress (gclc, gstp1, sod1 and cat), apoptosis (casp3a, casp6, casp8, casp9, aifm1 and tp53) and cell proliferation (pcna) related-genes were evaluated at 8 and 26 hpf. Caspase (3 and 9) activity was also determined at both time-points by colorimetric methods. Superoxide dismutase (SOD), catalase (CAT), glutathione levels (GSSG), caspase-9 and reactive oxygen species (ROS) were shown to be affected by ketamine exposure while in vivo analysis showed no difference in ROS. A significant up-regulation of superoxide dismutase (sod1) and catalase (cat) genes expression was also perceived. Ketamine-induced apoptosis was observed in vivo and confirmed by the apoptotic-related genes up-regulation. The overall results suggest that ketamine induced oxidative stress and apoptosis through the involvement of p53-dependent pathways in zebrafish embryos which could be important for the evaluation of the overall risk of ketamine in aquatic environments.

Determinación de fitoconstituyentes y evaluación de la actividad catártica en el extracto hidroalcohólico de Euphorbia huachanhana (huachangana) / Determination of phytoconstituents and evaluation of cathartic activity of Euphorbia huachanhana (Klotzsch & Garcke) Boiss (huachangana) hydroalcoholic extract

Objetivo. Determinar los fitoconstituyentes del extracto hidroalcohólico de Euphorbia huachanhana (Klotzsch & Garcke) Boiss (huachangana) y evaluar su actividad catártica. Materiales y métodos. Se elaboró un extracto hidroalcohólico con etanol al 70% (70:30) del tubérculo fresco de Euphorbia huachanhana (Klotzsch & Garcke) Boiss (huachangana), el cual fue analizado mediante un perfil fitoquímico cualitativo y el uso de espectroscopia UV/V y espectroscopia infrarroja (IR). Se tuvieron 36 ratones albinos, distribuidos en seis grupos de seis cada uno: Grupo I, suero fisiológico 0,9%); Grupo II, control positivo (senósido 250 mg/kg), y Grupos III, IV, V y VI (experimentales) a los que se les administró el extracto hidroalcohólico de Euphorbia huachanhana (Klotzsch & Garcke) Boiss (EH) a dosis 200 mg/kg, 400 mg/kg, 600 mg/kg y 800 mg/kg. Se evaluó la actividad catártica mediante la frecuencia de defecación y el porcentaje de motilidad intestinal con administración de carbón activado al 10%. Resultados. Se evidenció la presencia de flavonoides, compuestos fenólicos, carbohidratos, taninos, esteroides, azúcares reductores, aminoácidos y antraquinonas. Mediante espectroscopia UV/V se tuvo una banda de absorción de 290 nm lo que corresponde a una antraquinona; por espectroscopia infrarroja (IR) se evidenció la presencia de grupos hidroxilo y carbonilo. La mayor frecuencia de defecación (7,7 por hora) y el mejor tránsito intestinal (92,3%) fueron encontrados en el grupo que recibió EH a 200 mg/kg, estas diferencias fueron significativas en comparación al grupo blanco (p<0,05). Conclusiones. Se encontró, como molécula de interés, la presencia de una antraquinona con grupos funcionales hidroxilo y carbonilo; asimismo, la administración del extracto hidroalcohólico de Euphorbia huachanhana (Klotzsch & Garcke) Boiss a dosis de 200 mg/kg ofreció los mejores resultados en actividad catártica. Sin embargo, el diseño utilizado no permite establecer que la presencia de estas moléculas son la causa directa del efecto catártico observado.

Objective. To determine the phytoconstituents of the Euphorbia huachanhana (Klotzsch & Garcke) Boiss (huachangana) tuber hydroalcoholic extract and to evaluate its cathartic activity. Materials and methods. An hydroalcoholic extract was elaborated with 70% ethanol (70:30) using fresh tuber of Euphorbia huachanhana (Klotzsch & Garcke) Boiss (huachangana). That extract was analyzed for its phytochemical profile, and by UV/V spectroscopy and infrared spectroscopy (IR) to search some bioactive molecules. Thirty-six albino mice were distributed in six groups of six each one: Group I, 0.9% saline solution; Group II, Positive Control (Sennoside 250 mg/kg); and Groups III, IV, V and VI (Experimental) to which hydroalcoholic extract of Euphorbia huachanhana (Klotzsch & Garcke) Boiss (EH) were administered at doses 200 mg/kg, 400 mg/kg; 600 mg/kg and 800 mg/kg. The cathartic activity was evaluated by frequency of defecation and percentage of intestinal motility with administration of 10% activated carbon. Results. Presence of flavonoids, phenolic compounds, carbohydrates, tannins, steroids, reducing sugars, amino acids and anthraquinones was evidenced. An absorption band of 290 nm was observed by UV / V spectroscopy and infrared spectroscopy (IR), which corresponds to an anthraquinone, and the presence of hydroxyl and carbonyl groups was evidenced. The highest frequency of defecation (7.7 per hour) and the best intestinal transit (92.3%) were found on group which received EH at 200 mg/kg, these differences were statistically significant in comparison to the 0.9% saline solution group (p<0.05). Conclusions. The presence of an anthraquinone with hydroxyl and carbonyl groups was found as a molecule of interest. Additionally, the administration of Euphorbia huachanhana (Klotzsch & Garcke) Boiss hydroalcoholic extract at a dose of 200 mg / kg gave the best cathartic activity. However, this experimental design does not allow establishing that the presence of these molecules are the direct cause of the observed cathartic effect.

Apoptosis-related genes induced in response to ketamine during early life stages of zebrafish.

Increasing evidence supports that ketamine, a widely used anaesthetic, potentiates apoptosis during development through the mitochondrial pathway of apoptosis. Defects in the apoptotic machinery can cause or contribute to the developmental abnormalities previously described in ketamine-exposed zebrafish. The involvement of the apoptotic machinery in ketamine-induced teratogenicity was addressed by assessing the apoptotic signals at 8 and 24 hpf following 20min exposure to ketamine at three stages of early zebrafish embryo development (256 cell, 50% epiboly and 1-4 somites stages). Exposure at the 256-cell stage to ketamine induced an up-regulation of casp8 and pcna at 8 hpf while changes in pcna at the mRNA level were observed at 24 hpf. After the 50% epiboly stage exposure, the mRNA levels of casp9 were increased at 8 and 24 hpf while aifm1 was affected at 24 hpf. Both tp53 and pcna expressions were increased at 8 hpf. After exposure during the 1-4 somites stage, no meaningful changes on transcript levels were observed. The distribution of apoptotic cells and the caspase-like enzymatic activities of caspase-3 and -9 were not affected by ketamine exposure. It is proposed that ketamine exposure at the 256-cell stage induced a cooperative mechanism between proliferation and cellular death while following exposure at the 50% epiboly, a p53-dependent and -independent caspase activation may occur. Finally, at the 1-4 somites stage, the defence mechanisms are already fully in place to protect against ketamine-insult. Thus, ketamine teratogenicity seems to be dependent on the functional mechanisms present in each developmental stage.

Efecto de la ingestión de Banisteriopsis caapi y Psychotria viridis 'Binomio ayahuasca' en el hipocampo del cerebro de ratas / Effect of Banisteriopsis caapi and Psychotria viridis "Binomio ayahuasca" ingestion in the rat hippocampus

Objetivo: Evaluar el efecto de la ingestión de Banisteriopsis caapi y Psychotria viridis 'Binomio ayahuasca' en el hipocampo de cerebro de ratas. Diseño: Estudio experimental, descriptivo, analítico, transversal. Institución: Instituto de Investigación en Ciencias Farmacéuticas y Recursos Naturales, Facultad de Farmacia y Bioquímica, Universidad Nacional Mayor de San Marcos, Lima, Perú. Material biológico: Ratas. Intervenciones: Administración del binomio ayahuasca por vía orogástrica a ratas albinas adultas de la especie Rattus novergicus y de cepa Holtzman. A siete grupos de cinco con un peso promedio de 240 ± 30 g se les administró (GI) blanco, (GII) 2,5 mL diazepam, (GIII) 0,7 mL solución de Banisteriopsis caapi, (GIV) 0,7 mL Psychotria viridis, y a los grupos (GV), (GVI) y (GVII) se administró 0,7 mL, 3,5 mL y 7,0 mL de solución del binomio ayahuasca, respectivamente. Principales medidas de resultados: Macroscópicos: comportamiento de reflejos y actividad motora. Microscópicos: número de células piramidales y granulosas, y desorganización celular. Resultados: En el tamizaje fitoquímico del extracto se caracterizó presencia de alcaloides, antraquinonas, triterpenoides y esteroides, fenoles, flavonoides y saponinas. Los volúmenes de droga administradas a los grupos de intervención con el binomio en la dosis de 0,7 mL manifestaron significativo aumento en el número de células granulosas sobre las células piramidales; a dosis de 3,5 mL el número de células granulosas fue menor con presencia de células piramidales grandes y pequeñas; y a dosis de 7,0 mL se manifestó desorganización celular, presencia de células piramidales grandes y pequeñas, y aumento de células granulosas. Conclusiones: El extracto alcohólico del binomio ayahuasca presenta efecto neuropatológico en el hipocampo del cerebro de ratas. (AU)

Mitochondrial and liver oxidative stress alterations induced by N-butyl-N-(4-hydroxybutyl)nitrosamine: relevance for hepatotoxicity.

The most significant toxicological effect of nitrosamines like N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) is their carcinogenic activity, which may result from exposure to a single large dose or from chronic exposure to relatively small doses. However, its effects on mitochondrial liver bioenergetics were never investigated. Liver is the principal organ responsible for BBN metabolic activation, and mitochondria have a central function in cellular energy production, participating in multiple metabolic pathways. Therefore any negative effect on mitochondrial function may affect cell viability. In the present work, ICR male mice were given 0.05% of BBN in drinking water for a period of 12 weeks and were sacrificed one week later. Mitochondrial physiology was characterized in BBN- and control-treated mice. Transmembrane electric potential developed by mitochondria was significantly affected when pyruvate-malate was used, with an increase in state 4 respiration observed for pyruvate-malate (46%) and succinate (38%). A decrease in the contents of one subunit of mitochondrial complex I and in one subunit of mitochondrial complex IV was also observed. In addition, the activity of both complexes I and II was also decreased by BBN treatment. The treatment with BBN increases the susceptibility of liver mitochondria to the opening of the mitochondrial permeability transition pore. This susceptibility could be related with the increase in the production of H2 O2 by mitochondria and increased oxidative stress confirmed by augmented susceptibility to lipid peroxidation. These results lead to the conclusion that hepatic mitochondria are one primary target for BBN toxic action during liver metabolism.

Mitochondrial toxicity of the phyotochemicals daphnetoxin and daphnoretin--relevance for possible anti-cancer application.

Daphnetoxin is a daphnane type orthoester diterpene found exclusively in plants of the family Thymelaeaceae while daphnoretin, a bis-coumarin derivative that is the major constituent of the bark of some plants of this family, can also be found in Leguminosae and Rutaceae. These two compounds are recognized to have different biological effects, including a possible anti-cancer activity. The subject of the present research was to compare their mitochondrial toxicity and also investigate a possible selectivity towards tumor cell lines. Wistar rat liver mitochondria and three distinct cell lines were used to investigate compound-induced toxicity. The results indicate that both test compounds are toxic to isolated mitochondrial fractions, especially when used at concentrations higher than 100 microM. However, daphnetoxin presented the highest toxicity including increased proton leak in the inner mitochondrial membrane, increased induction of the mitochondrial permeability transition pore, inhibition of ATP synthase and inhibition of the mitochondrial respiratory chain. Both compounds also inhibited cell proliferation, regardless of the cell line used. Up to the maximal concentration tested in cells, no mitochondrial effects were detected by vital epifluorescence imaging, indicating that inhibition of cell proliferation may also originate from mitochondrial-independent mechanisms. The results warrant careful assessment of toxicity vs. pharmacology benefits of both molecules.

POLVO SEDIMENTABLE, ASMA BRONQUIAL Y ENFERMEDADES RESPIRATORIAS AGUDAS: San Antonio de los baños, 1994-2003 / San ntonio de loos baños. la habana / Settable dust, bronchial asthma and acute respiratory diseases

Las enfermedades respiratorias como el asma y las Infecciones Respiratorias Agudas (IRA), constituyen un importante problema de salud en el municipio San Antonio de los Baños. En esta investigación, se emplearon 238 resultados de muestras de polvo sedimentable, correspondiente a 120 meses de observación en un período de 10 años (1994-2003), procesadas en el Centro Municipal de Higiene y Epidemiología. Los datos de atenciones por IRA y crisis de asma se obtuvieron de la Dirección Municipal de Salud. Los resultados fundamentales consisten en que 75 % de la vigilancia de polvo sedimentable, estuvo por encima de la norma establecida (0.5 mg/cm²/30 días). A nivel municipal se reportan anualmente en promedio 29 176 casos notificados de IRA; así como 9 471 crisis de asma y se ubica dentro de los municipios que poseen alto riesgo por IRA. El análisis de la correlación y la tendencia por polvo sedimentable y las atenciones de IRA por 100 000 habitantes, reflejó que los picos de los mayores reportes por polvo sedimentable, anteceden a los mayores reportes de atenciones. Por otra parte, la comparación entre el polvo sedimentable y las tasas de atenciones por crisis de asma bronquial por 100 000 habitantes, arrojó que los picos de máxima contaminación por polvo sedimentable coinciden generalmente con mínimos de atenciones por crisis de asma bronquial. Todo ello sugiere que las crisis que ocurren ulteriormente, pueden ser asociadas con el polvo aún en suspensión. La condición de atmósfera cargada con concentraciones variables de polvo sedimentable, por encima de la norma estable-cida de 0.5 mg/cm²/30 días, pudiera exacerbar estas enfermedades respiratorias objeto de nuestro estudio.

Bronchial asthma (BA) and Acute Respiratory Infections (ARI) are an important health problem in the world as well as in San Antonio de los Baños municipality. This research used the 238 samples of pollution air from 1994 to 2003 that were correlated with chronological series of BA and ARI from the same period.The main results were that 75% of the samples of contamination surveillance system were over the normal levels of pollution.The annual average ARI were 29 176 attentions and 9471 attentions on BA. AB monthly attentions rate were in a range of nearly 2000 at 9000 / 100000 inhabitants and between 22 700 and 34 000 attentions on ARI. Atmospheric pollution, measured by particulate air pollution above the standard 0.5 mg/cm ²/30 days it was increased between May to August. The biggest reports by air pollution proceeded to ARI attention increased. BA was increased generally on March and April but when a pollution air decreased. The high level of pollution air may be an environment condition to increase the respiratory problems. Further studies are necessary to contribute in this field.

El envejeciente de Guarabo: un perfil biosicosocial del envejeciente en una comunidad de Puerto Rico / The aging person in Gurabo: a bio-psychosocial profile of the aging person in a community in Puerto Rico

The Department of Family Medicine of the University of Puerto Rico Medicine School performed a census of the inhabitants 60 years and older in the community of Gurabo. A total of 1,890 elderly were interviewed 51% were female and 49% male. Sixty percent were married and twenty-five percent were widowed. Eighty two percent still lived within a family setting and only seventeen percent lived alone 86% owned their own home despite ninety percent reporting incomes below $400.00 per month. Ninety five percent have access to medical services but 63% receive services in multiples sites. Eighteen percent were functionally impaired, fourteen percent intelectually impaired and five percent had impairment in both areas. Preliminary data show that the latter group had an annual mortality rate of 15% compared to only 3.6% and 1.3% respectively in the other two group. We concluded that: 1) functional and mental impairment are indicators of increased mortality. 2) although access to health care is not a problem this health care is episodic and interrupted being provided in multiple different sites

Comportamiento de la mortalidad por suicidio en la República Dominicana durante el período 1982-1988 / Behaviour of mortality by suicide in Republica Dominicana during 1982-1988

Mediante un estudio retrospectivo por recolección de información acerca del comportamiento del suicidio en el país, fueron analizadas las fichas policiales que registran el evento para el período 1982-1988. Los datos de 1988 se presentan incompletos. La presentación de este estudio se hizo durante el mes de junio de 1988. El objetivo consistió en dar seguimiento a las fichas policiales consideradas como la mejor fuente secundaria que registró el evento, para compararla con un estudio similar citado en el texto. Los hallazgos parecen seguir la misma tendencia que el estudio que refiere el período 1977 - 1988. Sin embargo, se connotan variaciones significativas en relación a los métodos utilizados y al aumento de las cifras absolutas por el sexo masculino. Se insiste en dar seguimiento a esta investigación y en ofrecer sugerencias al Departamento de Cartografía y Estadística de la Policía paa mejorar el dato que se capta y procesa